title: The role of Cε2, Cε3, and Cε4 domains in human and canine IgE and their contribution to FcεRIα interaction.
abstract: The Cε2 and Cε4 domains are considered as scaffolds, allowing Cε3 domains to assume an appropriate orientation to interact with FcεRI (Wurzburg and Jardetzky, 2002; Hunter et al., 2008). Human/canine IgE chimeric antibodies were expressed to assess the nature of the contribution of Cε2 and Cε4 domains to bind to and induce target cell degranulation via FcεRIα. Our results indicate that for (1) Cε3 domains in IgE of canine and human origin are the only necessary region for binding to FcεRIα. (2) The interaction of canine IgE with human sFcεRIα is significantly enhanced by contributions from both Cε2 and Cε4 domains of dog origin. (3) The canine/human IgE chimeric antibody construct rapidly dissociates from its the receptor when the canine Cε2 and Cε4 domains are replaced by the homologous human Fc domains which do not confer a conformation on the Cε3 domain to facilitate stable interaction with canine FcRIα. Kinetic constants for the binding of this chimera to the soluble extracellular domain of the receptor indicate an approximate 120-fold decrease in the affinity for canine sFcεRIα (ka=5.30 × 10(2)M(-1)s(-1)) and a 330-fold increase in the dissociation from canine sFcεRIα (KD=6.9 × 10(-6)M(-1)), compared to the wild type IgE kinetic constants (Ka=6.30 × 10(4)M(-1)s(-1); KD=2.1 × 10(-8)M(-1)). Although canine IgE does engage human FcεRIα, canine Cε2 and Cε4 do not contribute to the high-affinity of interaction with human FcεRIα. Upon replacement of human Cε2 and Cε4 domain by the canine homologues, human IgE Cε3 only retains a low affinity for the human receptor, which shows that Cε2 and Cε4 domains in human IgE Fc contribute significantly to the interaction with its cognate receptor.
authors.affiliation: Krebs Institute for Biomolecular Research, Department of Molecular Biology and Biotechnology, University of Sheffield, Firth Court, Western Bank, Sheffield S10 2TN, United Kingdom. Electronic address: mbp06hy@yahoo.co.uk.
authors.forename: Hongtu
authors.initials: H
authors.lastname: Ye
authors.forename: Jonathan E M
authors.initials: JE
authors.lastname: Housden
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Hunter
authors.forename: Sari
authors.initials: S
authors.lastname: Sabban
authors.forename: Birgit A
authors.initials: BA
authors.lastname: Helm
mesh_terms: D000818:Animals; D000915:Antibody Affinity; D001665:Binding Sites / Q000276:immunology; D004285:Dogs; D006801:Humans; D007073:Immunoglobulin E / Q000276:immunology* / Q000378:metabolism; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D051381:Rats; D017455:Receptors, IgE / Q000276:immunology / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000276:immunology
chemical_list: C543355:FcepsilonRI alpha-chain, human; D017455:Receptors, IgE; D011993:Recombinant Fusion Proteins; D007073:Immunoglobulin E
keywords: Human/canine Cɛ3 Fc chimeric IgE; Human/canine high-affinity receptor alpha chain; ca; canine IgE/FcɛRIα; hu; human IgE/FcɛRIα; s; soluble FcɛRIα